Li Ninghui, Zhao Ming, Liu Tengfei, Dong Lidong, Cheng Qun, Wu Junjiang, Wang Le, Chen Xi, Zhang Chuanzhong, Lu Wencheng, Xu Pengfei, Zhang Shuzhen
Key Laboratory of Soybean Biology of Chinese Education Ministry, Soybean Research Institute, Northeast Agricultural UniversityHarbin, China.
Jiamusi Branch of Heilongjiang Academy of Agricultural SciencesJiamusi, China.
Front Plant Sci. 2017 Jul 4;8:1185. doi: 10.3389/fpls.2017.01185. eCollection 2017.
Phytophthora root and stem rot caused by the oomycete pathogen is a destructive disease of soybean worldwide. Plant dirigent proteins (DIR) are proposed to have roles in biosynthesis of either lignan or lignin-like molecules, and are important for defense responses, secondary metabolism, and pathogen resistance. In the present work, a novel gene expressed sequence tag is identified as up-regulated in the highly resistant soybean cultivar 'Suinong 10' inoculated with . The full length cDNA is isolated using rapid amplification of cDNA ends, and designated (GenBank accession no. HQ_993047). The full length is 789 bp and contains a 567 bp open reading frame encoding a polypeptide of 188 amino acids. The sequence analysis indicated that GmDIR22 contains a conserved dirigent domain at amino acid residues 43-187. The quantitative real-time reverse transcription PCR demonstrated that soybean mRNA is expressed most highly in stems, followed by roots and leaves. The treatments with stresses demonstrated that is significantly induced by and gibberellic acid (GA), and also responds to salicylic acid, methyl jasmonic acid, and abscisic acid. The GmDIR22 is targeted to the cytomembrane when transiently expressed in Arabidopsis protoplasts. Moreover, The GmDIR22 recombinant protein purified from could effectively direct -coniferyl alcohol coupling into lignan (+)-pinoresinol. Accordingly, the overexpression of in transgenic soybean increased total lignan accumulation. Moreover, the lignan extracts from transgenic plants effectively inhibits hyphal growth. Furthermore, the transgenic overexpression of in the susceptible soybean cultivar 'Dongnong 50' enhances its resistance to . Collectively, these data suggested that the primary role of GmDIR22 is probably involved in the regulation of lignan biosynthesis, and which contributes to resistance to .
由卵菌病原体引起的疫霉根腐病和茎腐病是全球大豆的一种毁灭性病害。植物定向蛋白(DIR)被认为在木脂素或类木质素分子的生物合成中起作用,并且对防御反应、次生代谢和病原体抗性很重要。在本研究中,一个新的基因表达序列标签被鉴定为在接种了[病原体名称未给出]的高抗大豆品种‘绥农10’中上调表达。使用cDNA末端快速扩增技术分离出全长cDNA,并命名为GmDIR22(GenBank登录号:HQ_993047)。GmDIR22全长789 bp,包含一个567 bp的开放阅读框,编码一个188个氨基酸的多肽。序列分析表明,GmDIR22在氨基酸残基43 - 187处含有一个保守的定向结构域。定量实时逆转录PCR表明,大豆GmDIR22 mRNA在茎中表达最高,其次是根和叶。胁迫处理表明,GmDIR22受到[胁迫因素未给出]和赤霉素(GA)的显著诱导,并且对水杨酸、茉莉酸甲酯和脱落酸也有反应。当在拟南芥原生质体中瞬时表达时,GmDIR22定位于细胞膜。此外,从[表达系统未给出]中纯化的GmDIR22重组蛋白可以有效地将(+)-松柏醇偶联成木脂素(+)-松脂醇。因此,在转基因大豆中过表达GmDIR22增加了总木脂素积累量。此外,来自GmDIR22转基因植物的木脂素提取物有效地抑制了[病原体名称未给出]的菌丝生长。此外,在感病大豆品种‘东农50’中过表达GmDIR22增强了其对[病原体名称未给出]的抗性。总的来说,这些数据表明GmDIR22的主要作用可能参与木脂素生物合成的调控,并有助于对[病原体名称未给出]的抗性。